Quantitative profile of lipid classes in blood by normal phase chromatography with evaporative light scattering detector: application in the detection of lipid class abnormalities in liver cirrhosis

Clin Chim Acta. 2013 Jun 5:421:132-9. doi: 10.1016/j.cca.2013.02.007. Epub 2013 Feb 26.

Abstract

Background: The lack of analytical methods specific for each lipid class, particularly for phospholipids and sphyngolipids, makes necessary their separation by preparative techniques before quantification. LC-MS would be the election method but for daily work in the clinical laboratory this is not feasible for different reasons, both economic and time consuming. In the present work, we have optimized an HPLC method to quantify lipid classes in plasma and erythrocytes and applied it to samples from patients with cirrhosis.

Methods: Lipid classes were analyzed by normal phase liquid chromatography with evaporative light scattering detection. We employed a quaternary solvent system to separate twelve lipid classes in 15 min.

Results: Interday, intraday and recovery for quantification of lipid classes in plasma were excellent with our methodology. The total plasma lipid content of cirrhotic patients vs control subjects was decreased with diminished CE (81±33 vs 160±17 mg/dL) and PC (37±16 vs 60±19 mg/dL). The composition of erythrocytes showed a decrease in acidic phospholipids: PE, PI and PS.

Conclusion: Present methodology provides a reliable quantification of lipid classes in blood. The lipid profile of cirrhotics showed alterations in the PC/PE plasma ratio and in the phospholipid content of erythrocytes, which might reflect alterations in hepatocyte and erythrocyte membrane integrity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calibration
  • Chromatography, High Pressure Liquid / methods*
  • Erythrocytes / chemistry
  • Fatty Acids, Nonesterified / blood*
  • Fatty Acids, Nonesterified / classification
  • Humans
  • Light
  • Liver / metabolism
  • Liver / pathology
  • Liver Cirrhosis / blood*
  • Liver Cirrhosis / pathology
  • Phospholipids / blood*
  • Phospholipids / classification
  • Scattering, Radiation
  • Sensitivity and Specificity
  • Triglycerides / blood*
  • Triglycerides / classification

Substances

  • Fatty Acids, Nonesterified
  • Phospholipids
  • Triglycerides